Rao Chenglong, Mao Chan, Xia Yupei, Zhang Meijuan, Hu Zhiqiang, Yuan Siqi, Yang Wenbo, Yan Jingmin, Deng Ling, Cai Xiaolian, Mao Xuhu, Li Qian, Liao Yaling
Department of Clinical Microbiology and Immunology, College of Pharmacy and Medical Laboratory, Army Medical University (Third Military Medical University), Chongqing, China.
Department of Pharmacy, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, China.
Front Genet. 2020 Dec 8;11:585203. doi: 10.3389/fgene.2020.585203. eCollection 2020.
is a zoonotic pathogen that usually affects patients' lungs and causes serious melioidosis. The interaction of with its hosts is complex, and cellular response to infection in humans still remains to be elucidated. In this study, transcriptomic profiling of -infected human lung epithelial A549 cells was performed to characterize the cellular response dynamics during the early infection (EI) stage. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed by using the online databases DAVID 6.8 and KOBAS 3.0. Real-time quantitative PCR and western blot were used for validation experiments. Compared with the negative control group (NC), a set of 36 common genes varied over time with a cut-off level of 1.5-fold change, and a -value < 0.05 was identified. Bioinformatics analysis indicated that the PERK-mediated unfolded protein response (UPR) was enriched as the most noteworthy biological process category, which was enriched as a branch of UPR in the signaling pathway of protein processing in the endoplasmic reticulum. Other categories, such as inflammatory responses, cell migration, and apoptosis, were also focused. The molecular chaperone Bip (GRP78), PERK, and PERK sensor-dependent phosphorylation of eIF2α (p-eIF2α) and ATF4 were verified to be increasing over time during the EI stage, suggesting that infection activated the PERK-mediated UPR in A549 cells. Collectively, these results provide important initial insights into the intimate interaction between and lung epithelial cells, which can be further explored toward the elucidation of the cellular mechanisms of infections in humans.
是一种人畜共患病原体,通常会影响患者肺部并引发严重的类鼻疽病。它与宿主的相互作用很复杂,人类对其感染的细胞反应仍有待阐明。在本研究中,对感染该病原体的人肺上皮A549细胞进行了转录组分析,以表征早期感染(EI)阶段的细胞反应动态。使用在线数据库DAVID 6.8和KOBAS 3.0进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)通路分析。采用实时定量PCR和蛋白质印迹法进行验证实验。与阴性对照组(NC)相比,一组36个常见基因随时间变化,变化阈值为1.5倍,且P值<0.05。生物信息学分析表明,PERK介导的未折叠蛋白反应(UPR)作为最值得注意的生物学过程类别得到富集,它在内质网蛋白质加工信号通路中作为UPR的一个分支得到富集。其他类别,如炎症反应、细胞迁移和细胞凋亡,也受到关注。分子伴侣Bip(GRP78)、PERK以及eIF2α(p-eIF2α)和ATF4的PERK传感器依赖性磷酸化在EI阶段随时间增加得到验证,这表明该病原体感染激活了A549细胞中PERK介导的UPR。总的来说,这些结果为该病原体与肺上皮细胞之间的密切相互作用提供了重要的初步见解,可进一步探索以阐明人类感染该病原体的细胞机制。